Elastase/ELANE/ELA2 BioAssay(TM) ELISA Kit (Mouse)

Neutrophil elastase is a serine protease of neutrophil and monocyte granules. Its key physiologic role is in innate host defense, but it can also participate in tissue remodeling and possesses secretagogue actions important to local inflammatory responses. Elastases form a subfamily of serine proteases that hydrolyze many proteins in addition to elastin. Humans have six elastase genes which encode structurally similar proteins. The encoded preproprotein is proteolytically processed to generate the active protease. Following activation, this protease hydrolyzes proteins within specialized neutrophil lysosomes, called azurophil granules, as well as proteins of the extracellular matrix. This protein also degrades the outer membrane protein A (OmpA) of E. coli as well as the virulence factors of such bacteria as Shigella, Salmonella and Yersinia. Mutations in this gene are associated with cyclic neutropenia and severe congenital neutropenia (SCN). This gene is present in a gene cluster on chromosome 19. Sample Type: Cell lysates, cell culture supernates, serum, plasma (EDTA or Heparin). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Mouse Elastase/ELANE/ELA2. Sensitivity: <10pg/ml, Range: 31.2pg/ml-2000pg/ml, Specificity: Recognizes natural and recombinant Mouse Elastase/ELANE/ELA2. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Mouse Elastase/ELANE/ELA2 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A specific for Elastase/ELANE/ELA2 has been precoated onto 96-well plates. Standards(Expression system for standard: NSO, Immunogen sequence: S27-R260) and test samples are added to the wells, a biotinylated detection specific for Elastase/ELANE/ELA2 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Mouse Elastase/ELANE/ELA2 amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Mouse Elane antibody, 1x plate, Lyophilized recombinant Mouse Elane standard, 2x10ng/tube, Biotinylated anti-Mouse Elane antibody, 1x130ul (dilution 1:100), Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100), Sample diluent buffer, 1x30ml, Antibody diluent buffer, 1x12ml, ABC diluent buffer, 1x12ml, TMB color developing agent, 1x10ml, TMB stop solution, 1x10ml, PBS washing buffer Powder for 1000ml, 1x buffer, Storage and Stability: Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.